[Pharmaceutical intervention in duration of antimicrobial treatment at hospital ambit]. / Actuación farmacéutica en el control de la duración del tratamiento con antimicrobianos en el ámbito hospitalario.

OBJECTIVE: To estimate the acceptance of the pharmaceutical intervention in controlling duration of antimicrobial therapy and to evaluate their impact on optimizing the treatment. METHODS: Prospective observational study for two years in a General University Hospital. For the patients record, we followed non critical adult patients with antibiotic treatment. When the duration of antimicrobial treatment not complied with established criteria for each antibiotic and pathology, there was a communication with the physician, at which is recommended to assess the need for continue treatment. The acceptance of pharmaceutical intervention was collected and afterwards we analyzed the impact of this work by antimicrobial consumption and incidence of Clostridium difficile. RESULTS: . In 122 patients the pharmacist made a pharmaceutical intervention due to prolonged antibiotic treatment. The most prevalent antibiotics were ß-lactams, specifically meropenem. The intravenous administration was more frequent. In 77 cases it was decided to recommend the suspension of treatment, we conducted an orally prospective intervention at 70.15 % and the rest of interventions were written. Acceptance was 65.95 % and 65.00%, respectively. During the study period, the DDD of the antimicrobials decreased by 8.89% and expenditure on antimicrobials one 40.12%. The incidence of C. difficile was stable. CONCLUSIONS: . In a hospital, a pharmaceutical counselling program on the duration of antimicrobial therapy is well accepted by the prescriber physician, but it must be improved. The route of information does not affect the degree of acceptance. These actions could involve a reduction of antimicrobial consumption.

Activation-associated phenotype of CD3 T cells in acute graft-versus-host disease.

During the effector phase of graft-versus-host disease (GvHD) response, donor T cells play an essential role and they are believed to change the expression of activation and co-stimulatory markers associated with functional alloreactivity. We analysed the expression of CD25, CD69, HLA-DR, CD154 and CD134 on CD4+ and CD8+ T cells by flow cytometry during acute GvHD (aGvHD) in 24 patients receiving human leucocyte antigen (HLA)-identical stem cell transplants. Expression of these molecules in nine patients with stages I-IV aGvHD was compared with 15 patients without aGvHD (n = 15). Serial analysis showed that peripheral blood of aGvHD patients presented a significant increase of CD4+ CD25+ cells (P < 0.03), CD4+ CD69+ (P < 0.04) and CD4+ CD134+ cells (P < 0.01). Additionally, there was a significant increase in CD8+ cells expressing CD134 (P = 0.007) and CD154 (P = 0.02). After resolution of aGvHD, the increased expression of these molecules returned to values comparable to patients without aGvHD. Only two of the 15 patients without clinical signs of aGvHD presented activated T cells that could not be attributed to development of aGvHD. In summary, our data show that multiple activation molecules are preferentially up-regulated on CD4+ and CD8+ T cells from patients with aGvHD. These patients had a significant increase in the expression of the co-stimulatory molecules CD134 and CD154.

DRB1*1446 allele variations in the group-specific annealing sites of common amplification primers.

A cord blood unit from the Umbilical Cord Bank of Barcelona was initially typed by sequence-based typing (SBT) as DRB1* 0701. However, the subsequent confirmation by sequence-specific oligonucleotide probes (SSOP) revealed an unusual hybridization pattern that did not fit any of the known HLA-DRB1 alleles or allelic combinations. Molecular cloning and sequencing with primers located at introns 1 and 2 provided the definitive identification of a novel DRB1*1446 allele. It was similar to DRB1*1402 at exon 2 except for nucleotide substitutions at codons 10, 11 and 12 (from YST to LRK) that coincide with the 5'-end group specific site for the annealing of common amplification primers used by SBT and sequence-specific primers.

Sequence of the HLA-B*1573 allele.

We report here the sequence of an HLA B*15 allele, B*1573. Initially, a cord blood unit from the Umbilical Cord Bank of Barcelona was typed by sequence-specific oligonucleotide hybridization and revealed an unusual hybridization pattern. After the cloning, the sequence-based typing assigned two different alleles: B*07021, and a second allele identical to B*1558 in exons 2 and 3, except for a single nucleotide substitution in exon 3, which changed codon 140 from Phe to Leu (TTX-->TTA).

Molecular analysis of HLA-DRB1 polymorphism in north-east Spain.

The DRB1* polymorphism in 941 randomly selected individuals from the Umbilical Cord Blood Bank of Barcelona (92.75% of Spanish origin) was determined by sequence-based typing. The HLA profile was similar to that of other Mediterranean populations, with DRB1*0701 and *0301 being the most frequent alleles. This may be a consequence of the mixture of alleles as a result of migration from contiguous geographical areas.

Identification and sequencing of the HLA-B*4106 allele.

We report a novel HLA-B*41 allele (HLA-B*4106) initially detected by an unusual sequence-specific oligonucleotide (SSO) hybridization pattern and identified by sequence-based HLA typing. Molecular cloning and sequencing determined that the new HLA-B allele was identical to the HLA-B*4101 in exon 2 and 3 except for a single nucleotide substitution in the exon 3 changing codon 204 from Glu to Gln (GAG-->CAG). In addition, intron 2 was identical to the published HLA-B*4104 intron 2, except for the base 509 (C-->G).

Synthesis and anti-inflammatory evaluation of new sulfamoylheterocarboxylic derivatives.

A series of new sulfamoylthiophene and sulfamoylpyrazole carboxylic acid derivatives was synthesized. Some of these compounds show interesting analgesic properties and significant nonsteroidal anti-inflammatory activities in several models of inflammation.

Treatment with an anti-CD4 monoclonal antibody strongly ameliorates established rat adjuvant arthritis.

Some experimental arthritic diseases can be prevented by treatment with anti-CD4 MoAbs. Trials with ongoing disease have not been successful so far. The aim of this study was to ascertain whether W3/25 could reverse adjuvant arthritis (AA), when beginning treatment on day 14, i.e. when the disease was established. Moreover, one group of animals treated with the anti-CD4 MoAb received OX8 MoAb at the same time, thus depleting CD8+ cells from circulation. During treatment with W3/25, a strong amelioration of inflammatory signals were observed, as assessed by means of paw volume increase and arthritic score. However, when treatment stopped, a rebound to arthritis signals occurred. The parallel depletion of CD8+ cells did not modify these effects, thus the combined treatment W3/25 + OX8 gave the same amelioration as treatment with W3/25 alone. These findings indicate that CD4+ cells play an important role in perpetuating rat AA. Moreover, CD8+ cells do not seem to have a regulatory role int he CD4+ cells responsible for the inflammatory response.

Comparison of four lymphocyte isolation methods applied to rodent T cell subpopulations and B cells.

The aim of this study was to establish the validity of four lymphocyte isolation methods. The effects of three different erythrocyte lysing methods commonly used in the analysis of human cells, namely, lysis by ammonium chloride (AC), Becton Dickinson lysis (BDL) and the Coulter Q-Prep (CQP) preparation system were established by flow cytometry on rat lymphocyte subsets. The results were compared with those obtained with a Ficoll-Isopaque (FI) density gradient procedure adapted for use with rat cells. Lymphocyte isolation by AC or FI gradient was performed before labelling the lymphocyte subpopulations, whereas the BDL and CQP methods were performed after staining the cells in whole blood. The FI gradient yielded the lowest CD5+, CD4+ and CD25+ cell percentages. On the other hand AC lysis produced higher percentages of T cells and lower percentages of B cells than the other methods studied. The percentages obtained after BDL or CQP methods for T lymphocyte subsets and B cells were found to be reproducible. The commercial methods (BDL and CQP) are faster but rather expensive, whereas AC lysis and FI gradient separations are cheap and particularly useful when there is a requirement to culture the cells.

Administration of a nondepleting anti-CD4 monoclonal antibody (W3/25) prevents adjuvant arthritis, even upon rechallenge: parallel administration of a depleting anti-CD8 monoclonal antibody (OX8) does not modify the effect of W3/25.

The aim of this study was to determine the effects of the anti-CD4 monoclonal antibody (mAb) W3/25, found to be nondepleting, on the onset of rat adjuvant arthritis (AA), and, in addition, to ascertain whether depletion of CD8+ cells during the same period could interfere with those effects. Female Wistar rats in which AA had been induced were treated with W3/25 and/or OX8 (anti-rat CD8) mAb during the latency period of arthritis. W3/25 alone or in combination with OX8 prevented the inflammatory process of AA. When the protected groups were rechallenged with a second dose of Mycobacterium butyricum no arthritis was observed. Protected and nonprotected arthritic animals developed the same anti-mycobacteria antibody levels as the arthritic control group. This study indicates that a nondepleting anti-CD4 mAb can prevent AA, while CD8+ lymphocytes do not appear relevant for the development of AA and do not seem to have a regulatory role for CD4+ cells.

In vitro and in vivo effects of three bismuth compounds on fermentation by colonic bacteria.

Three bismuth compounds (tripotassium dicitrate bismuthate, bismuth subsalicylate, and bismuth subnitrate) were tested in vitro and in vivo for their effect on fermentation by colonic bacteria. The studies in vitro were done with use of a technique designed to determine the effect of each one of the bismuth compounds on the fermentation of several stool samples that had been mixed with lactose as additional fermentable substrate (fermentation of lactose-enriched stools, FLES). The three bismuth compounds reduced FLES significantly in 47 (81%) of 58 of the stool samples used to test their effect. Bismuth subsalicylate, which reduced FLES in 10 of 10 stool samples, showed the greatest reduction (mean reduction, 74%; P less than .0001). The in vivo studies, done in six flatulent patients, showed significant reduction (P less than .01) of colonic fermentation of ingested raffinose by oral bismuth subnitrate given for 8 days.

On the rate of tubal pregnancy contralateral to the corpus luteum.

Ectopic implantation of the embryo in the tube opposite to the ovary containing the corpus luteum constitutes evidence of peritoneal or uterine transmigration of the egg. The frequency of this phenomenon was reinvestigated utilizing histopathologic confirmation of the side of the corpus luteum. A tubal pregnancy contralateral to the ovulating ovary was found in 28% of 67 cases, indicating that either the oocyte, the zygote or the embryo had entered the tube in which implantation took place from a medial site such as the peritoneal or the uterine cavity rather than directly from the ovulating ovary. Assuming that once in the medial site there is equal chance of entering either tube, it follows that in 56% of tubal pregnancies the egg has entered the tube from a midline location. Attempts to recover the oocyte from the tubes in normal women were successful in fewer than 5% of cases contralateral to the corpus luteum. It is concluded that tubal pregnancy is associated with a significant increase in the occurrence of transmigration of the egg.

[Myasthenia and sleep]. / Myasthénie et sommeil.

A Central Nervous System (CNS) acetylcholine receptors involvement in Myasthenia Gravis (MG) has been suggested but never fully demonstrated. On the other hand, although the question concerning the neurotransmitters connected to REM sleep is still unsettled, the importance of acetylcholine as a neurotransmitter involved in the maintenance of this sleep stage has been outlined. The spontaneous night sleep organization has been studied in 9 male subjects with MG and results are compared with those obtained in normal subjects matched for age and sex. Statistically significant differences have been found: 1) slow-waves sleep better represented in MG patients; 2) average REM period length shorter in MG patients; 3) instability of the EEG consisting in a tendency toward lightening of sleep. These findings may confirm the presence of a disturbance in the CNS cholinergic activity of MG patients.

Sleep and human cerebellar pathology.

The organization and the quantitative evaluation of sleep stages were studied in patients with cerebellar syndrome. 25 polygraphic night recordings were scored according to the criteria suggested by Rechtschaffen & Kales (1968) and the sleep parameters investigated were referred to those indicated by Williams et al. (1974). The results matched with a homogeneous sample of normal subjects. Significant alterations were observed for several parameters of sleep, and the possible causes discussed.